Hydroxyalkyl substituted pyrido-7-pyrimidin-7-ones

ABSTRACT

Compounds of the Formula: 
                         
where X 1 , Ar 1 , R 1 , and R 2  are as defined herein, and compositions comprising the same. Also provided are methods for using compounds of Formula I in treating p38 mediated disorders in a patient.

CROSS REFERENCE TO RELATED APPLICATIONS

This Application is a continuation of U.S. patent application Ser. No.10/987,656 filed on Nov. 12, 2004, now U.S. Pat. No. 7,348,331, whichclaims the benefit under Title 35 U.S.C. 119(e) of U.S. ProvisionalApplication No. 60/519,789 filed Nov. 13, 2003. The disclosures of theaforementioned applications are incorporated herein by reference intheir entirety.

FIELD OF THE INVENTION

The present invention relates to pyridopyrimidines and derivativesthereof. In particular, the present invention provides 2,6-disubstituted7-oxo-pyrido[2,3-d]pyrimidines, a process for their manufacture,pharmaceutical preparations comprising the same, and methods for usingthe same.

BACKGROUND OF THE INVENTION

Mitogen-activated protein kinases (MAP) is a family of proline-directedserine/threonine kinases that activate their substrates by dualphosphorylation. The kinases are activated by a variety of signalsincluding nutritional and osmotic stress, UV light, growth factors,endotoxin and inflammatory cytokines. One group of MAP kinases is thep38 kinase group that includes various isoforms (e.g., p38α, p39β, p38γand p38δ). The p38 kinases are responsible for phosphorylating andactivating transcription factors as well as other kinases, and areactivated by physical and chemical stress, pro-inflammatory cytokinesand bacterial lipopolysaccharide.

More importantly, the products of the p38 phosphorylation have beenshown to mediate the production of inflammatory cytokines, including TNFand IL-1, and cyclooxygenase-2. Each of these cytokines has beenimplicated in numerous disease states and conditions. For example, TNF-αis a cytokine produced primarily by activated monocytes and macrophages.Its excessive or unregulated production has been implicated as playing acausative role in the pathogenesis of rheumatoid arthritis. Morerecently, inhibition of TNF production has been shown to have broadapplication in the treatment of inflammation, inflammatory boweldisease, multiple sclerosis and asthma.

TNF has also been implicated in viral infections, such as HIV, influenzavirus, and herpes virus including herpes simplex virus type-1 (HSV-1),herpes simplex virus type-2 (HSV-2), cytomegalovirus (CMV),varicella-zoster virus (VZV), Epstein-Barr virus, human herpes virus-6(HHV-6), human herpesvirus-7 (HHV-7), human herpesvirus-8 (HHV-8),pseudorabies and rhinotracheitis, among others.

Similarly, IL-1 is produced by activated monocytes and macrophages, andplays a role in many pathophysiological responses including rheumatoidarthritis, fever and reduction of bone resorption.

Additionally, the involvement of p38 has been implicated in stroke,Alzheimer's disease, osteoarthritis, lung injury, septic shock,angiogenesis, dermatitis, psoriasis and atopic dermatitis. J. Exp. Opin.Ther. Patents, 2000, 10(1).

The inhibition of these cytokines by inhibition of the p38 kinase is ofbenefit in controlling, reducing and alleviating many of these diseasestates.

Certain 6-aryl-pyrido[2,3-d]pyrimidin-7-ones, -7-imines and 7-thionesare disclosed as inhibitors of protein tyrosine kinase mediated cellularproliferation in WO 96/34867, published Nov. 7, 1996 (Warner Lambert).Other 6-aryl-pyrido[2,3-d]pyrimidines and naphthyridines are alsodisclosed as inhibitors of tyrosine kinase in WO 96/15128, published May23, 1996 (Warner Lambert). 6-alkyl-pyrido[2,3-d]pyrimidin-7-ones aredisclosed as inhibitors of cyclin-dependent kinases in WO 98/33798,published Aug. 6, 1998 (Warner Lambert). Certain4-amino-pyridopyrimidines are disclosed as inhibitors of dihydrofolatereductase in EP 0 278 686A1, published Aug. 8, 1988 (WellcomeFoundation).

SUMMARY OF THE INVENTION

One aspect of the present invention provides compounds of the formula:

where

-   -   X¹ is O, S(O)_(n) (where n is 0, 1 or 2), or C═O;    -   Ar¹ is aryl or heteroaryl;    -   R¹ is alkoxyalkyl, alkyl, cycloalkyl, cycloalkylalkyl,        heterocyclyl, hydroxyalkyl, or hydroxycycloalkyl; and    -   R² is hydroxyalkyl, oxoalkyl or hydroxycycloalkyl.

Another aspect of the present invention provides a pharmaceuticalformulation comprising a Compound of Formula I and a pharmaceuticallyacceptable carrier, diluent, or excipient therefor.

While certain substituted pyrido-7-pyrimidin-7-ones are known to beactive in an enzymatic in vitro assay against p38 (see for example,commonly assigned U.S. patent application Ser. No. 10/073,845, filedFeb. 11, 2002, entitled “6-Substituted Pyrido-Pyrimidines”, which isincorporated herein by reference in its entirety), surprisingly andunexpectedly, the present inventor has discovered that Compound ofFormula I have a significantly higher activity in a lipopolysacchararide(LPS) induced human whole blood cysteine production assay than compoundsthat have previously been disclosed.

Compounds of Formula I are inhibitors of protein kinases, and exhibiteffective activity against p38 in vivo. They are selective against p38kinase relative to cyclin-dependent kinases and tyrosine kinases.Therefore, compounds of the present invention can be used for thetreatment of diseases mediated by the pro-inflammatory cytokines such asTNF and IL-1. Thus another aspect of the present invention provides amethod for treating p38 mediated diseases or conditions in which atherapeutically effective amount of a Compound of Formula I isadministered to a patient.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

Unless otherwise stated, the following terms used in the specificationand claims have the meanings given below:

“Alkoxyalkyl” means a moiety of the formula R^(a)—O—R^(b)—, where R^(a)is alkyl and R^(b) is alkylene as defined herein. Exemplary alkoxyalkylgroups include, by way of example, 2-methoxyethyl, 3-methoxypropyl,1-methyl-2-methoxyethyl, 1-(2-methoxyethyl)-3-methoxypropyl, and1-(2-methoxyethyl)-3-methoxypropyl.

“Alkyl” means a linear saturated monovalent hydrocarbon radical of oneto six carbon atoms or a branched saturated monovalent hydrocarbonradical of three to six carbon atoms, e.g., methyl, ethyl, propyl,2-propyl, n-butyl, iso-butyl, tert-butyl, pentyl, and the like.

“Alkylene” means a linear saturated divalent hydrocarbon radical of oneto six carbon atoms or a branched saturated divalent hydrocarbon radicalof three to six carbon atoms, e.g., methylene, ethylene,2,2-dimethylethylene, propylene, 2-methylpropylene, butylene, pentylene,and the like.

“Aryl” means a monovalent monocyclic or bicyclic aromatic hydrocarbonradical which is optionally substituted independently with one or moresubstituents, preferably one, two or three, substituents preferablyselected from the group consisting of alkyl, hydroxy, alkoxy, haloalkyl,haloalkoxy, halo, nitro, cyano, amino, monoalkylamio, dialkylamino,methylenedioxy, ethylenedioxy and acyl. A particularly preferred arylsubstituent is halide. More specifically the term aryl includes, but isnot limited to, phenyl, chlorophenyl, fluorophenyl, difluorophenyl (suchas 2,4- and 2,6-difluorophenyl), methoxyphenyl, 1-naphthyl, 2-naphthyl,and the derivatives thereof.

“Cycloalkyl” refers to a saturated monovalent cyclic hydrocarbon radicalof three to seven ring carbons e.g., cyclopropyl, cyclobutyl,cyclohexyl, 4-methyl-cyclohexyl, and the like. Cycloalkyl may optionallybe substituted with one or more substituents, preferably one, two orthree, substituents. Preferably, cycloalkyl substituent is selected fromthe group consisting of alkyl, hydroxy, alkoxy, haloalkyl, haloalkoxy,halo, amino, monoalkylamio, dialkylamino, and acyl. A particularlypreferred group of cycloalkyl substituents include alkyl, hydroxy,alkoxy, haloalkyl, haloalkoxy, and halo. An especially preferred groupof cycloalkyl substituents include alkyl, hydroxy, alkoxy, and halo.

“Cycloalkylalkyl” refers to a moiety of the formula R^(c)-R^(d)—, whereR^(c) is cycloalkyl and R^(d) is alkylene as defined herein.

“Halo” and “halide” are used interchangeably herein and refer to fluoro,chloro, bromo, or iodo. Preferred halides are fluoro and chloro withfluoro being a particularly preferred halide.

“Haloalkyl” means alkyl substituted with one or more same or differenthalo atoms, e.g., —CH₂Cl, —CF₃, —CH₂CF₃, —CH₂CCl₃, and the like.

“Heteroaryl” means a monovalent monocyclic or bicyclic radical of 5 to12 ring atoms having at least one aromatic ring containing one, two, orthree ring heteroatoms selected from N, O, or S (preferably N or O), theremaining ring atoms being C, with the understanding that the attachmentpoint of the heteroaryl radical will be on an aromatic ring. Theheteroaryl ring is optionally substituted independently with one or moresubstituents, preferably one or two substituents, selected from alkyl,haloalkyl, hydroxy, alkoxy, halo, nitro or cyano. More specifically theterm heteroaryl includes, but is not limited to, pyridyl, furanyl,thienyl, thiazolyl, isothiazolyl, triazolyl, imidazolyl, isoxazolyl,pyrrolyl, pyrazolyl, pyrimidinyl, benzofuranyl, tetrahydrobenzofuranyl,isobenzofuranyl, benzothiazolyl, benzoisothiazolyl, benzotriazolyl,indolyl, isoindolyl, benzoxazolyl, quinolyl, tetrahydroquinolinyl,isoquinolyl, benzimidazolyl, benzisoxazolyl or benzothienyl,imidazo[1,2-a]-pyridinyl, imidazo[2,1-b]thiazolyl, and the derivativesthereof.

“Heterocyclyl” means a saturated or unsaturated non-aromatic cyclicradical of 3 to 8 ring atoms in which one or two ring atoms areheteroatoms selected from N, O, or S(O)_(n) (where n is an integer from0 to 2), preferably N or O, the remaining ring atoms being C, where oneor two C atoms may optionally be replaced by a carbonyl group. Theheterocyclyl ring may be optionally substituted independently with one,two, or three substituents selected from alkyl, haloalkyl, hydroxyalkyl,halo, nitro, cyano, cyanoalkyl, hydroxy, alkoxy, amino, monoalkylamino,dialkylamino, aralkyl, —(X)_(n)—C(O)R^(e) (where X is O or NR^(f), n is0 or 1, R^(e) is hydrogen (where X is NR^(f)), alkyl, haloalkyl, hydroxy(when n is 0), alkoxy, amino, monoalkylamino, dialkylamino, oroptionally substituted phenyl, and R^(f) is H or alkyl),alkylene-C(O)R^(g) (where R^(g) is alkyl, —OR^(h) or NR^(i)R^(j) andR^(b) is hydrogen, alkyl or haloalkyl, and R^(i) and R^(j) areindependently hydrogen or alkyl), or —S(O)_(n)R^(k) (where n is aninteger from 0 to 2) such that when n is 0, R^(k) is hydrogen, alkyl,cycloalkyl, or cycloalkylalkyl, and when n is 1 or 2, R^(k) is alkyl,cycloalkyl, cycloalkylalkyl, amino, acylamino, monoalkylamino, ordialkylamino. A particularly preferred group of heterocyclylsubstituents include alkyl, haloalkyl, hydroxyalkyl, halo, hydroxy,alkoxy, amino, monoalkylamino, dialkylamino, aralkyl, and—S(O)_(n)R^(k). In particular, the term heterocyclyl includes, but isnot limited to, tetrahydrofuranyl, pyridinyl, tetrahydropyranyl,piperidino, N-methylpiperidin-3-yl, piperazino, N-methylpyrrolidin-3-yl,3-pyrrolidino, morpholino, thiomorpholino, thiomorpholino-1-oxide,thiomorpholino-1,1-dioxide, 4-(1,1-dioxo-tetrahydro-2H-thiopyranyl),pyrrolinyl, imidazolinyl, N-methanesulfonyl-piperidin-4-yl, and thederivatives thereof, each of which may be optionally substituted.

“Hydroxyalkyl” means an alkyl moiety as defined herein, substituted withone or more, preferably one, two or three hydroxy groups, provided thatthe same carbon atom does not carry more than one hydroxy group.Representative examples include, but are not limited to, hydroxymethyl,2-hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl,1-(hydroxymethyl)-2-methylpropyl, 2-hydroxybutyl, 3-hydroxybutyl,4-hydroxybutyl, 2,3-dihydroxypropyl, 2-hydroxy-1-hydroxymethylethyl,2,3-dihydroxybutyl, 3,4-dihydroxybutyl and2-(hydroxymethyl)-3-hydroxypropyl

“Hydroxycycloalkyl” means a cycloalkyl moiety as defined herein whereinone, two or three hydrogen atoms in the cycloalkyl radical have beenreplaced with a hydroxy substituent. Representative examples include,but are not limited to; 2-, 3-, or 4-hydroxycyclohexyl, and the like.

“Leaving group” has the meaning conventionally associated with it insynthetic organic chemistry, i.e., an atom or a group capable of beingdisplaced by a nucleophile and includes halo (such as chloro, bromo, andiodo), alkanesulfonyloxy, arenesulfonyloxy, alkylcarbonyloxy (e.g.,acetoxy), arylcarbonyloxy, mesyloxy, tosyloxy,trifluoromethanesulfonyloxy, aryloxy (e.g., 2,4-dinitrophenoxy),methoxy, N,O-dimethylhydroxylamino, and the like.

“Optionally substituted phenyl” means a phenyl ring which is optionallysubstituted independently with one or more substituents, preferably oneor two substituents selected from the group consisting of alkyl,hydroxy, alkoxy, haloalkyl, haloalkoxy, halo, nitro, cyano, amino,methylenedioxy, ethylenedioxy, and acyl.

“Oxoalkyl” means an alkyl group which is substituted with one or morecarbonyl oxygen moiety (i.e., ═O), such as a moiety of the formulaR^(z)—C(═O)R^(y)—, wherein R^(y) is alkylene and R^(z) is alkyl.Exemplary oxoalkyl groups include 2-propanon-3-yl,2-methyl-3-butanon-4-yl, and the like.

“Pharmaceutically acceptable excipient” means an excipient that isuseful in preparing a pharmaceutical composition that is generally safe,non-toxic and neither biologically nor otherwise undesirable, andincludes excipient that is acceptable for veterinary use as well ashuman pharmaceutical use. A “pharmaceutically acceptable excipient” asused in the specification and claims includes both one and more than onesuch excipient.

“Pharmaceutically acceptable salt” of a compound means a salt that ispharmaceutically acceptable and that possesses the desiredpharmacological activity of the parent compound. Such salts include: (1)acid addition salts, formed with inorganic acids such as hydrochloricacid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, andthe like; or formed with organic acids such as acetic acid, propionicacid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvicacid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid,fumaric acid, tartaric acid, citric acid, benzoic acid,3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic-acid,methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid,2-hydroxyethanesulfonic acid, benzenesulfonic acid,4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid,4-toluenesulfonic acid, camphorsulfonic acid,4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid,3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid,lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoicacid, salicylic acid, stearic acid, muconic acid, and the like; or (2)salts formed when an acidic proton present in the parent compound eitheris replaced by a metal ion, e.g., an alkali metal ion, an alkaline earthion, or an aluminum ion; or coordinates with an organic base such asethanolamine, diethanolamine, triethanolamine, tromethamine,N-methylglucamine, and the like.

The terms “pro-drug” and “prodrug” are used interchangeably herein andrefer to any compound which releases an active parent drug according toFormula I in vivo when such prodrug is administered to a mammaliansubject. Prodrugs of a compound of Formula I are prepared by modifyingone or more functional group(s) present in the compound of Formula I insuch a way that the modification(s) may be cleaved in vivo to releasethe parent compound. Prodrugs include compounds of Formula I wherein ahydroxy, amino, sulfhydryl, carboxy or carbonyl group in a compound ofFormula I is bonded to any group that may be cleaved in vivo toregenerate the free hydroxyl, amino, or sulfhydryl group, respectively.Examples of prodrugs include, but are not limited to, esters (e.g.,acetate, dialkylaminoacetates, formates, phosphates, sulfates, andbenzoate derivatives) and carbamates (e.g., N,N-dimethylaminocarbonyl)of hydroxy functional groups, esters groups (e.g. ethyl esters,morpholinoethanol esters) of carboxyl functional groups, N-acylderivatives (e.g. N-acetyl) N-Mannich bases, Schiff bases and enaminonesof amino functional groups, oximes, acetals, ketals and enol esters ofketone and aldehyde functional groups in compounds of Formula I, and thelike, See Bundegaard, H. “Design of Prodrugs” p 1-92, Elesevier,N.Y.-Oxford (1985).

“Protecting group” refers to a grouping of atoms that when attached to areactive group in a molecule masks, reduces or prevents that reactivity.Examples of protecting groups can be found in T. W. Green and P. G.Futs, Protective Groups in Organic Chemistry, (Wiley, 2 d ed. 1991) andHarrison and Harrison et al., Compendium of Synthetic Organic Methods,Vols. 1-8 (John Wiley and Sons, 1971-1996). Representative aminoprotecting groups include, formyl, acetyl, trifluoroacetyl, benzyl, benzyloxycarbonyl (CBZ), tert-butoxycarbonyl (Boc), trimethyl silyl (TMS),2-trimethylsilyl-ethanesulfonyl (SES), trityl and substituted tritylgroups, allyloxycarbonyl, 9-fluorenylmethyloxycarbonyl (FMOC),nitro-veratryloxycarbonyl (NVOC), and the like. Representative hydroxyprotecting groups include those where the hydroxy group is eitheracylated or alkylated such as benzyl, and trityl ethers as well as alkylethers, tetrahydropyranyl ethers, trialkylsilyl ethers and allyl ethers.

“Treating” or “treatment” of a disease includes: (1) preventing thedisease, i.e., causing the clinical symptoms of the disease not todevelop in a mammal that may be exposed to or predisposed to the diseasebut does not yet experience or display symptoms of the disease; (2)inhibiting the disease, i.e., arresting or reducing the development ofthe disease or its clinical symptoms; or (3) relieving the disease,i.e., causing regression of the disease or its clinical symptoms.

“A therapeutically effective amount” means the amount of a compoundthat, when administered to a mammal for treating a disease, issufficient to effect such treatment for the disease. The“therapeutically effective amount” will vary depending on the compound,the disease and its severity and the age, weight, etc., of the mammal tobe treated.

The term “treating”, “contacting” or “reacting” when referring to achemical reaction, means to add or mix two or more reagents underappropriate conditions to produce the indicated and/or the desiredproduct. It should be appreciated that the reaction which produces theindicated and/or the desired product may not necessarily result directlyfrom the combination of two reagents which were initially added, i.e.,there can be one or more intermediates which are produced in the mixturewhich ultimately leads to the formation of the indicated and/or thedesired product.

Compounds of the Invention

One embodiment of the present invention provides a compound of theformula:

where

-   -   X¹ is O, S(O)_(n) (where n is 0, 1 or 2), or C═O;    -   Ar¹ is aryl or heteroaryl;    -   R¹ is alkoxyalkyl, alkyl, cycloalkyl, cycloalkylalkyl,        heterocyclyl, hydroxyalkyl, or hydroxycycloalkyl; and    -   R² is hydroxyalkyl, oxoalkyl pr hydroxycycloalkyl.

The compounds of the present invention can exist in unsolvated forms aswell as solvated forms, including hydrated forms. In general, thesolvated forms, including hydrated forms, are equivalent to unsolvatedforms and are intended to be encompassed within the scope of the presentinvention. In addition to the compounds described above, the compoundsof the present invention include all tautomeric forms. Furthermore, thepresent invention also includes all pharmaceutically acceptable salts ofthose compounds along with prodrug forms of the compounds and allstereoisomers whether in a pure chiral form or a racemic mixture orother form of mixture.

The compounds of Formula I are capable of further formingpharmaceutically acceptable acid addition salts. All of these forms arewithin the scope of the present invention.

Pharmaceutically acceptable acid addition salts of the compounds ofFormula I include salts derived from inorganic acids such ashydrochloric, nitric, phosphoric, sulfuric, hydrobromic, hydriodic,phosphorous, and the like, as well as the salts derived from organicacids, such as aliphatic mono- and dicarboxylic acids,phenyl-substituted alkanoic acids, hydroxy alkanoic acids, alkanedioicacids, aromatic acids, aliphatic and aromatic sulfonic acids, etc. Suchsalts thus include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite,nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate,metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate,propionate, caprylate, isobutyrate, oxalate, malonate, succinate,suberate, sebacate, fumarate, maleate, mandelate, benzoate,chlorobenzoate, methylbenzoate, dinitrobenzoate, phthalate,benzenesulfonate, toluenesulfonate, phenylacetate, citrate, lactate,maleate, tartrate, methanesulfonate, and the like. Also contemplated aresalts of amino acids such as arginate and the like and gluconate,galacturonate (see, for example, Berge S. M., et al., “PharmaceuticalSalts,” J. of Pharmaceutical Science, 1977, 66, 1-19).

The acid addition salts of the basic compounds can be prepared bycontacting the free base form with a sufficient amount of the desiredacid to produce the salt in the conventional manner. The free base formcan be regenerated by contacting the salt form with a base and isolatingthe free base in the conventional manner. The free base forms differfrom their respective salt forms somewhat in certain physical propertiessuch as solubility in polar solvents, but otherwise the salts areequivalent to their respective free base for purposes of the presentinvention.

In one embodiment, Ar¹ is aryl. A particularly preferred Ar¹ isoptionally substituted phenyl. In certain embodiments, Ar¹ is phenyloptionally substituted one or more times with alkyl, halo, haloalkyl oralkoxy. A more preferred Ar¹ is disubstituted phenyl such as2,4-disubstituted phenyl. Still more preferably, Ar¹ is 2,4-dihalosubstituted phenyl. An especially preferred Ar¹ is 2,4-difluorophenyl.

Still in another embodiment, X¹ is O.

In another embodiment, R¹ is alkoxyalkyl, alkyl, cycloalkyl,cycloalkylalkyl, hydroxyalkyl, or heterocyclyl. Within this group, aparticularly preferred R¹ includes optionally substitutedtetrahydropyranyl, 1-methyl-2-methoxyethyl, optionally substitutedcyclopentyl, optionally substituted cyclopropyl, iso-propyl, optionallysubstituted cyclohexyl, 1-(2-hydroxyethyl)-3-hydroxypropyl,1-hydroxymethyl-2-hydroxypropyl, 1-hydroxymethyl-3-hydroxypropyl,1-methylpropyl, 2-hydroxy-1-methylethyl,1-(2-methoxyethyl)-3-methoxypropyl, N-methanesulfonyl piperidinyl,ethyl, methyl, 2-hydroxypropyl, neopentyl, 1,1-dimethyl-2-hydroxyethyl,1-(hydroxymethyl)propyl, 2-methylpropyl, cyclopropylmethyl, optionallysubstituted cyclobutyl, 1,2-dimethyl-2-hydroxypropyl, and1-(hydroxymethyl)-2-hydroxyethyl.

Yet in another embodiment, a preferred R¹ is hydroxyalkyl, with2-hydroxy-1-methylethyl being a particularly preferred R¹. Especiallypreferred R¹ includes enantiomerically enriched 2-hydroxy-1-methylethyl,i.e., (R)- and (S)-2-hydroxy-1-methylethyl.

In one specific embodiment, R² is 2-hydroxyethyl, 3-hydroxypropyl,2-hydroxypropyl, 1-(2-hydroxyethyl)-3-hydroxypropyl, or 2-oxopropyl.

In another embodiment, R² is hydroxyalkyl. Within this group, aparticularly preferred R² is 2-hydroxyethyl, 3-hydroxypropyl,2-hydroxypropyl, and 1-(2-hydroxyethyl)-3-hydroxypropyl. An especiallypreferred R² is 2-hydroxypropyl.

Still further, combinations of the preferred groups described hereinform other preferred embodiments. For example, in one particularlypreferred embodiment R¹ is (R)- or (S)-2-hydroxy-1-methylethyl, R² is(R)- or (S)-2-hydroxypropyl, or 2-oxopropyl, X¹ is O and Ar¹ is2,4-difluorophenyl.

In one embodiment the present invention provides a compound of formula Iwherein Ar¹ is aryl and X¹ is O. In another embodiment the presentinvention provides a compound of formula I wherein Ar¹ is aryl, X¹ is Oand R¹ is alkoxyalkyl, alkyl, cycloalkyl, cycloalkylalkyl, hydroxyalkyl,or heterocyclyl. In still another embodiment the present inventionprovides a compound of formula I wherein Ar¹ is aryl, X¹ is O and R¹ istetrahydropyranyl, 1-methyl-2-methoxyethyl, cyclopentyl, cyclopropyl,iso-propyl, cyclohexyl, 1-(2-hydroxyethyl)-3-hydroxypropyl,1-hydroxymethyl-2-hydroxypropyl, 1-hydroxymethyl-3-hydroxypropyl,1-methylpropyl, 2-hydroxy-1-methylethyl,1-(2-methoxyethyl)-3-methoxypropyl, N-methanesulfonyl piperidinyl,ethyl, methyl, 2-hydroxypropyl, neopentyl, 1,1-dimethyl-2-hydroxyethyl,1-(hydroxymethyl)propyl, 2-methylpropyl, cyclopropylmethyl, cyclobutyl,1,2-dimethyl-2-hydroxypropyl, or 1-(hydroxymethyl)-2-hydroxyethyl. Inyet another embodiment the present invention provides a compound offormula I wherein Ar¹ is aryl, X¹ is O, R¹ is tetrahydropyranyl,1-methyl-2-methoxyethyl, cyclopentyl, cyclopropyl, iso-propyl,cyclohexyl, 1-(2-hydroxyethyl)-3-hydroxypropyl,1-hydroxymethyl-2-hydroxypropyl, 1-hydroxymethyl-3-hydroxypropyl,1-methylpropyl, 2-hydroxy-1-methylethyl,1-(2-methoxyethyl)-3-methoxypropyl, N-methanesulfonyl piperidinyl,ethyl, methyl, 2-hydroxypropyl, neopentyl, 1,1-dimethyl-2-hydroxyethyl,1-(hydroxymethyl)propyl, 2-methylpropyl, cyclopropylmethyl, cyclobutyl,1,2-dimethyl-2-hydroxypropyl, or 1-(hydroxymethyl)-2-hydroxyethyl and R²is 2-hydroxyethyl, 3-hydroxypropyl, 2-hydroxypropyl,1-(2-hydroxyethyl)-3-hydroxypropyl, or 2-oxopropyl.

In one embodiment the present invention provides a compound of formula Iwherein Ar¹ is aryl, X¹ is O, R¹ is (R)-2-hydroxy-1-methylethyl or(S)-2-hydroxy-1-methylethyl and R² is 2-oxopropyl, (R)-2-hydroxypropylor (S)-2-hydroxypropyl.

In one embodiment the present invention provides a compound of formula Iwherein Ar¹ is aryl, X¹ is O and R¹ is hydroxyalkyl. In anotherembodiment the present invention provides a compound of formula Iwherein Ar¹ is aryl, X¹ is O, R¹ is hydroxyalkyl and R² is hydroxyalkyl.

In still another embodiment the present invention provides a compound offormula I wherein Ar¹ is aryl, X¹ is O, R¹ is hydroxyalkyl and R² is2-hydroxyethyl, 3-hydroxypropyl, 2-hydroxypropyl, or1-(2-hydroxyethyl)-3-hydroxypropyl.

In another embodiment the present invention provides a compound offormula I wherein Ar¹ is aryl, X¹ is O;

-   -   R¹ is (R)-2-hydroxy-1-methylethyl and R² is (R)-2-hydroxypropyl;    -   R¹ is (R)-2-hydroxy-1-methylethyl and R² is (S)-2-hydroxypropyl;    -   R¹ is (S)-2-hydroxy-1-methylethyl and R² is (R)-2-hydroxypropyl;        or    -   R¹ is (S)-2-hydroxy-1-methylethyl and R² is (S)-2-hydroxypropyl.

In one embodiment the present invention provides a compound of formula Iwherein R¹ is hydroxyalkyl. In another embodiment the present inventionprovides a compound of formula I wherein R¹ is hydroxyalkyl and R² ishydroxyalkyl. In still another embodiment the present invention providesa compound of formula I wherein R¹ is hydroxyalkyl, R² is hydroxyalkyland Ar¹ is aryl.

In one embodiment the present invention provides a compound of formula Iwherein R² is hydroxyalkyl.

In certain embodiments, the compounds of the invention may be of theformula:

wherein:

-   -   m is from 0 to 4;    -   each R³ is alkyl, halo, alkoxy or haloalkyl; and    -   R¹ and R² are as described herein.

In specific embodiments, m is 1 and R³ is halo.

In still other embodiments, m is 2 and R³ is halo.

In one embodiment the present invention provides a compound of formulaII wherein R¹ is alkoxyalkyl, alkyl, cycloalkyl, cycloalkylalkyl,hydroxyalkyl, or heterocyclyl.

In one embodiment the present invention provides a compound of formulaII wherein R¹ is tetrahydropyranyl, 1-methyl-2-methoxyethyl,cyclopentyl, cyclopropyl, iso-propyl, cyclohexyl,1-(2-hydroxyethyl)-3-hydroxypropyl, 1-hydroxymethyl-2-hydroxypropyl,1-hydroxymethyl-3-hydroxypropyl, 1-methylpropyl,2-hydroxy-1-methylethyl, 1-(2-methoxyethyl)-3-methoxypropyl,N-methanesulfonyl piperidinyl, ethyl, methyl, 2-hydroxypropyl,neopentyl, 1,1-dimethyl-2-hydroxyethyl, 1-(hydroxymethyl)propyl,2-methylpropyl, cyclopropylmethyl, cyclobutyl,1,2-dimethyl-2-hydroxypropyl, or 1-(hydroxymethyl)-2-hydroxyethyl. Inanother embodiment the present invention provides a compound of formulaII wherein R¹ is tetrahydropyranyl, 1-methyl-2-methoxyethyl,cyclopentyl, cyclopropyl, iso-propyl, cyclohexyl,1-(2-hydroxyethyl)-3-hydroxypropyl, 1-hydroxymethyl-2-hydroxypropyl,1-hydroxymethyl-3-hydroxypropyl, 1-methylpropyl,2-hydroxy-1-methylethyl, 1-(2-methoxyethyl)-3-methoxypropyl,N-methanesulfonyl piperidinyl, ethyl, methyl, 2-hydroxypropyl,neopentyl, 1,1-dimethyl-2-hydroxyethyl, 1-(hydroxymethyl)propyl,2-methylpropyl, cyclopropylmethyl, cyclobutyl,1,2-dimethyl-2-hydroxypropyl, or 1-(hydroxymethyl)-2-hydroxyethyl and R²is 2-hydroxyethyl, 3-hydroxypropyl, 2-hydroxypropyl,1-(2-hydroxyethyl)-3-hydroxypropyl, or 2-oxopropyl.

In one embodiment the present invention provides a compound of formulaII wherein R¹ is (R)-2-hydroxy-1-methylethyl or(S)-2-hydroxy-1-methylethyl and R² is 2-oxopropyl, (R)-2-hydroxypropylor (S)-2-hydroxypropyl.

In one embodiment the present invention provides a compound of formulaII wherein R¹ and R² are hydroxyalkyl.

In one embodiment the present invention provides a compound of formulaII wherein:

-   -   R¹ is (R)-2-hydroxy-1-methylethyl and R² is (R)-2-hydroxypropyl;    -   R¹ is (R)-2-hydroxy-1-methylethyl and R² is (S)-2-hydroxypropyl;    -   R¹ is (S)-2-hydroxy-1-methylethyl and R² is (R)-2-hydroxypropyl;        or    -   R¹ is (S)-2-hydroxy-1-methylethyl and R² is (S)-2-hydroxypropyl.

In another embodiment the present invention provides a compound offormula II wherein R¹ and R² are hydroxyalkyl, n is 1 and R³ is halo.

In another embodiment the present invention provides a compound offormula II wherein R¹ and R² are hydroxyalkyl, n is 2 and R³ is halo.

Representative compounds in accordance with the invention are shown inTable I.

TABLE 1 # Structure Name 1

6-(2,4-Difluoro-phenoxy)-8-(2-hydxoxy-ethyl)-2-(tetrahydro-pyran-4-ylamino)-8H-pyrido[2,3-d]pyrimidin- 7-one 2

6-(2,4-Difluoro-phenoxy)-8-(3-hydroxy-propyl)-2-(tetrahydro-pyran-4-ylamino)-8H-pyrido[2,3-d]pyrimidin- 7-one 3

6-(2,4-Difluoro-phenoxy)-8-(2-hydroxy-propyl)-2-(tetrahydro-pyran-4-ylamino)-8H-pyrido[2,3-d]pyrimidin- 7-one 4

6-(2,4-Difluoro-phenoxy)-8-[3-hydroxy-1-(2-hydroxy-ethyl)-propyl]-2-((S)-2-methoxy-1-methyl-ethylamino)-8H-pyrido[2,3-d]pyrimidin-7-one 5

2-Cyclopentylamino-6-(2,4-difluoro-phenoxy)-8-((S)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7-one 6

2-Cyclopropylamino-6-(2,4-difluoro-phenoxy)-8-((S)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7-one 7

6-(2,4-Difluoro-phenoxy)-8-((S)-2-hydroxy-propyl)-2-isopropylamino-8H-pyrido[2,3-d]pyrimidin-7-one 8

2-Cyclohexylamino-6-(2,4-difluoro-phenoxy)-8-((S)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7-one 9

6-(2,4-Difluoro-phenoxy)-2-[3-hydroxy-1-(2-hydroxy-ethyl)-propylamino]-8-((S)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7-one 10

6-(2,4-Difluoro-phenoxy)-8-((S)-2-hydroxy-propyl)-2-(tetrahydro-pyran-4-ylamino)-8H-pyrido[2,3-d]pyrimidin- 7-one 11

6-(2,4-Difluoro-phenoxy)-2-((1R,2R)-2-hydroxy-1-hydroxymethyl-propylamino)-8-((S)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7-one 12

6-(2,4-Difluoro-phenoxy)-2-((S)-3-hydroxy-1-hydroxymethyl-propylamino)-8-((S)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7-one 13

6-(2,4-Difluoro-phenoxy)-2-((R)-3-hydroxy-1-hydroxymethyl-propylamino)-8-((S)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7-one 14

2-((S)-sec-Butylamino)-6-(2,4-difluoro-phenoxy)-8-((S)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7-one 15

2-((R)-sec-Butylamino)-6-(2,4-difluoro-phenoxy)-8-((S)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrinaidin-7-one 16

6-(2,4-Difluoro-phenoxy)-2-((S)-2-hydroxy-1-methyl-ethylamino)-8-((S)-2-hydroxy-propyl)-8H-pyrido[2,3- d]pyrimidin-7-one 17

6-(2,4-Difluoro-phenoxy)-2-((R)-2-hydroxy-1-methyl-ethylamino)-8-((S)-2-hydroxy-propyl)-8H-pyrido[2,3- d]pyrimidin-7-one 18

6-(2,4-Difluoro-phenoxy)-8-((S)-2-hydroxy-propyl)-2-((S)-2-methoxy-1-methyl-ethylamino)-8H-pyrido[2,3- d]pyrimidin-7-one 19

6-(2,4-Difluoro-phenoxy)-8-((S)-2-hydroxy-propyl)-2-[3-methoxy-1-(2-methoxy-ethyl)-propylamino]-8H-pyrido[2,3-d]pyrimidin-7-one 20

6-(2,4-Difluoro-phenoxy)-8-((R)-2-hydroxy-propyl)-2-(tetrahydro-pyran-4-ylamino)-8H-pyrido[2,3-d]pyrimidin- 7-one 21

6-(2,4-Difluoro-phenoxy)-8-((S)-2-hydroxy-propyl)-2-(1-methanesulfonyl-piperidin-4-ylamino)-8H-pyrido[2,3- d]pyrimidin-7-one 22

2-Cyclopropylamino-6-(2,4-difluoro-phenoxy)-8-((R)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7-one 23

6-(2,4-Difluoro-phenoxy)-8-((R)-2-hydroxy-propyl)-2-isopropylamino-8H-pyrido[2,3-d]pyrimidin-7-one 24

6-(2,4-Difluoro-phenoxy)-2-ethylamino-8-((R)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7-one 25

6-(2,4-Difluoro-phenoxy)-8-((R)-2-hydroxy-propyl)-2-methylamino-8H-pyrido[2,3-d[pyrimidin-7-one 26

2-((S)-sec-Butylamino)-6-(2,4-difluoro-phenoxy)-8-((R)-2-hydroxy-propyl)-8H-pyrido[2,3-d[pyrimidin-7-one 27

2-((R)-sec-Butylamino)-6-(2,4-difluoro-phenoxy)-8-((R)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7-one 28

6-(2,4-Difluoro-phenoxy)-8-((S)-2-hydroxy-propyl)-2-((R)-2-hydroxy-propylamino)-8H-pyrido[2,3-d]pyrimidin- 7-one 29

6-(2,4-Difluoro-phenoxy)-8-((S)-2-hydroxy-propyl)-2-((S)-2-hydroxy-propylamino)-8H-pyrido[2,3-d]pyrimidin- 7-one 30

6-(2,4-Difluoro-phenoxy)-2-(2,2-dimethyl-propylamino)-8-((S)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7- one 31

6-(2,4-Difluoro-phenoxy)-2-(2-hydroxy-1,1-dimethyl-ethylamino)-8-((R)-2-hydroxy-propyl)-8H-pyrido[2,3- d]pyrimidin-7-one 32

6-(2,4-Difluoro-phenoxy)-2-((S)-1-hydroxymethyl-propylamino)-8-((S)-2-hydroxy-propyl)-8H-pyrido[2,3- d]pyrimidin-7-one33

6-(2,4-Difluoro-phenoxy)-2-((R)-1-hydroxymethyl-propylamino)-8-((S)-2-hydroxy-propyl)-8H-pyrido[2,3- d]pyrimidin-7-one34

6-(2,4-Difluoro-phenoxy)-8-((R)-2-hydroxy-propyl)-2-((S)-2-hydroxy-propylamino)-8H-pyrido[2,3-d]pyrimidin- 7-one 35

6-(2,4-Difluoro-phenoxy)-8-((R)-2-hydroxy-propyl)-2-((R)-2-hydroxy-propylamino)-8H-pyrido[2,3-d]pyrimidin- 7-one 36

6-(2,4-Difluoro-phenoxy)-2-(2-hydroxy-1,1-dimethyl-ethylamino)-8-((S)-2-hydroxy-propyl)-8H-pyrido[2,3- d]pyrimidin-7-one 37

6-(2,4-Difluoro-phenoxy)-2-((R)-1-hydroxymethyl-propylamino)-8-((R)-2-hydroxy-propyl)-8H-pyrido[2,3- d]pyrimidin-7-one38

6-(2,4-Difluoro-phenoxy)-8-((R)-2-hydroxy-propyl)-2-isobutylamino-8H-pyrido[2,3-d]pyrimidin-7-one 39

2-(Cyclopropylmethyl-amino)-6-(2,4-difluoro-phenoxy)-8-((R)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7- one 40

2-Cyclobutylamino-6-(2,4-difluoro-phenoxy)-8-((R)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7-one 41

6-(2,4-Difluoro-phenoxy)-2-((S)-2-hydroxy-1-methyl-ethylamino)-8-((R)-2-hydroxy-propyl)-8H-pyrido[2,3- d]pyrimidin-7-one 42

6-(2,4-Difluoro-phenoxy)-2-((R)-2-hydroxy-1-methyl-ethylamino)-8-((R)-2-hydroxy-propyl)-8H-pyrido[2,3- d]pyrimidin-7-one 43

6-(2,4-Difluoro-phenoxy)-2-(2,2-dimethyl-propylamino)-8-((R)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7- one 44

6-(2,4-Difluoro-phenoxy)-2-((S)-2-hydroxy-1,2-dimethyl-propylamino)-8-((R)-2-hydroxy-propyl)-8H-pyrido[2,3- d]pyrimidin-7-one45

6-(2,4-Difluoro-phenoxy)-2-((S)-2-hydroxy-1-methyl-ethylamino)-8-(2-oxo-propyl)-8H-pyrido[2,3-d]pyrimidin- 7-one 46

6-(2,4-Difluoro-phenoxy)-2-((R)-2-hydroxy-1-methyl-ethylamino)-8-(2-oxo-propyl)-8H-pyrido[2,3-d]pyrimidin- 7-one 47

6-(2,4-Difluoro-phenoxy)-2-(2-hydroxy-1-hydroxymethyl-ethylamino)-8-((S)-2-bydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7-one

While the forms of the invention herein constitute presently preferredembodiments, many others are possible. It is not intended herein tomention all of the possible equivalent forms or ramifications of theinvention. It is understood that the terms used herein are merelydescriptive rather than limiting, and that various changes can be madewithout departing from the spirit or scope of the invention.

Processes for Preparing the Compounds

The compounds of the present invention can be prepared by a variety ofmethods including by methods disclosed in commonly assigned U.S. patentapplication Ser. No. 10/073,845, which was previously incorporated byreference. In one aspect of the present invention, a method forpreparing compounds of Formula I is shown in Scheme 1 below. It shouldbe appreciated that although the scheme often indicates exactstructures, methods of the present invention apply widely to analogouscompounds of Formula I, given appropriate consideration to protectionand deprotection of reactive functional groups by methods standard tothe art of organic chemistry. For example, hydroxy groups, in order toprevent unwanted side reactions, sometimes need to be protected (e.g.,converted to ethers or esters) during chemical reactions at other sitesin the molecule. The hydroxy protecting group is then removed to providethe free hydroxy group. Similarly, amino groups and carboxylic acidgroups can be protected (e.g., by derivatization) to protect themagainst unwanted side reactions. Typical protecting groups, and methodsfor attaching and cleaving them, are described fully in the aboveincorporated references by T. W. Greene and P. G. M. Wuts, ProtectiveGroups in Organic Synthesis, 3^(rd) edition, John Wiley & Sons, NewYork, 1999, and Harrison and Harrison et al., Compendium of SyntheticOrganic Methods, Vols. 1-8 (John Wiley and Sons, 1971-1996).

Treatment of a compound of Formula Ia with a hydroxyalkyl amine (R²—NH₂)provides a compound of Formula Ib. This reaction is conveniently carriedout in a solvent which is inert under the reaction conditions,preferably a halogenated aliphatic hydrocarbon, especiallydichloromethane, an optionally halogenated aromatic hydrocarbon, or anopen-chain or cyclic ether such as tetrahydrofuran (THF), a formamide ora lower alkanol. Suitably, the reaction is carried out at about −20° C.to about 120° C., typically at about 0° C. Often a base, such astrialkyl amine, preferably triethylamine, is added to the reactionmixture.

Reduction of a compound of Formula Ib provides an alcohol of Formula Ic.This reduction is typically carried out using lithium aluminum hydridein a manner well known to those of skill in the art (e.g., in a solventthat is inert under the conditions of the reduction, preferably anopen-chain or cyclic ether, especially tetrahydrofuran, at about −20° C.to about 70° C., preferably at about 0° C. to about room temperature).

Oxidation of an alcohol of Formula Ic provides a carboxaldehyde ofFormula Id. The oxidation is typically carried out with manganesedioxide, although numerous other methods can also be employed (see, forexample, ADVANCED ORGANIC CHEMISTRY, 4^(TH) ED., March, John Wiley &Sons, New York (1992)). Depending on the oxidizing agent employed, thereaction is carried out conveniently in a solvent which is inert underthe specific oxidation conditions, preferably a halogenated aliphatichydrocarbon, especially dichloromethane, or an optionally halogenatedaromatic hydrocarbon. Suitably, the oxidation is carried out at about 0°C. to about 60° C.

Reaction of a carboxaldehyde of Formula Id with an ester,Ar¹—X¹CH₂—CO₂R′ (where R′ is an alkyl group, and Ar¹ and X¹ are thosedefined herein) in the presence of a base provides a compound of FormulaIe. Any relatively non-nucleophilic base can be used includingcarbonates, such as potassium carbonate, lithium carbonate, and sodiumcarbonate; bicarbonates, such as potassium bicarbonate, lithiumbicarbonate, and sodium bicarbonate; amines, such as secondary andtertiary amines; and resin bound amines such as 1,3,4,6,7,8-hexahydro-2Hpyrimido[1,2-a]pyrimidine. Conveniently, the reaction is carried out ina solvent which is relatively polar but inert under the reactionconditions, preferably an amide such as dimethyl formamide,N-substituted pyrrolidinone, especially 1-methyl-2-pyrrolidinone, and ata temperature of about 25° C. to about 150° C.

Oxidation of Ie with an oxidizing agent, e.g., a peracid such as3-chloroperbenzoic acid (i.e., MCPBA) or Oxone®, provides a sulfone (If)which can be converted to a variety of target compounds. Typically theoxidation of Ie is carried out in a solvent which is inert under theconditions of the oxidation. For example, when MCPBA is used as theoxidizing agent, the solvent is preferably a halogenated aliphatichydrocarbon, especially chloroform. When Oxone® is used as the oxidizingagent, the solvent is preferably methanol, aqueous ethanol or aqueoustetrahydrofuran. The reaction temperature depends on the solvent used.For an organic solvent, the reaction temperature is generally at about−20° C. to about 50° C., preferably about 0° C. to about roomtemperature. When water is used as the solvent, the reaction temperatureis generally from about 0° C. to about 50° C., preferably about 0° C. toabout room temperature. Alternatively, the oxidation may be carriedunder catalytic conditions with rhenium/peroxide based reagents, see(“Oxidation of Sulfoxides by Hydrogen Peroxide, Catalyzed byMethyltrioxorhenium(VII)”, Lahti et al., Inorg. Chem., 2000, 39,2164-2167; “Rhenium oxo complexes in catalytic oxidations, Catal. Today,2000, 55, 317-363, and “A Simple and Efficient Method for thePreparation of Pyridine N-Oxides”, Coperet et al., J. Org. Chem., 1998,63, 1740-1741.

Reacting the compound If with an amine (R¹—NH₂) provides the compoundsof Formula I. The reaction can be carried out in the presence or absenceof solvent. Conveniently, the reaction is carried out at temperatures offrom about 0° C. to about 200° C., more preferably about roomtemperature to about 150° C. Alternatively, in some cases rather thanusing the sulfone If, the sulfide Ie or the corresponding sulfoxide canbe reacted directly with an amine (R¹—NH₂) to provide the compounds ofFormula I.

One of skill in the art will understand that certain modifications tothe above schemes are contemplated and within the scope of the presentinvention. For example, certain steps will involve the use of protectinggroups for functional groups that are not compatible with particularreaction conditions.

Alternatively, compounds of Formula I can also be prepared by the methodshown in Scheme 2 below. While the reactions of Scheme 2 are shown interms of specific compounds, it will be readily apparent to thoseskilled in the art that the method of Scheme 2 may be used with all ofthe compounds of the invention.

As shown in Scheme 2, treatment of a diethyl acetal IIa with a thioureaprovides a pyrimidine compound IIb. This reaction is convenientlycarried out in an alcoholic solvent in the presence of a base, such assodium methoxide. Methylation of the thiol group, e.g., with methyliodide, then provides thioether IIc.

The thioether IIc may then be treated with an α-aryloxyester IId such asethyl (2,4-difluorophenoxy)acetate to afford a pyrido-pyrimidonethioether IIe. This reaction may be carried out, for example, by heatingin the presence sodium carbonate or other mild base inn-methylpyrrolidinone or other polar aprotic solvent.

Reaction of thioether IIe with propylene carbonate or like carbonate,under polar aprotic solvent conditions, to yield an N-hyrdoxyalkylpyrido-pyrimidone thioether IIf. This reaction may be facilitated byheating in the presence of potassium carbonate.

The thioether IIf is then oxidized to provide the correspondingpyrido-pyrimidone sulfone IIg. This oxidation may be carried out usinghydrogen peroxide in the presence of acetic acid in polar solvent suchas methylene dichloride. The oxidation may alternatively be carried outusing Oxone® or MCPBA in the manner described above for Scheme 1.

Treatment of sulfone IIg with a hydroxyamine, wherein the hydroxyl groupis suitably protected, affords a pyrido-pyrimidone compound IIh inaccordance with the invention. This reaction may be carried out withheating as described above with reference to Scheme 1.

Pyridopyrimidinone IIf can also be prepared by alkylatingpyridopyrimidinone IIe with an epoxide instead of a carbonate as shownin Scheme 3 below. The reaction of Scheme 3 may be carried out byheating compound IIe under pressure in the presence of excess propyleneoxide in N-methylpyrrolidinone or under other polar aprotic solventconditions.

Pharmaceutical Compositions Containing the Compounds

The compounds of Formula I can be used as medicaments, e.g., in the formof pharmaceutical preparations. The pharmaceutical preparations can beadministered enterally, e.g., orally in the form of tablets, coatedtablets, dragées, hard and soft gelatine capsules, solutions, emulsionsor suspensions, nasally, e.g., in the form of nasal sprays, or rectally,e.g., in the form of suppositories. However, they may also beadministered parenterally, e.g., in the form of injection solutions.

The compounds of Formula I can be processed with pharmaceutically inert,organic or inorganic carriers for the production of pharmaceuticalpreparations. Lactose, corn starch or derivatives thereof, talc, stearicacid or its salts and the like can be used, for example, as suchcarriers for tablets, coated tablets, dragées and hard gelatinecapsules. Suitable carriers for soft gelatine capsules are, for example,vegetable oils, waxes, fats, semi-solid and liquid polyols and the like;depending on the nature of the active ingredient no carriers are,however, usually required in the case of soft gelatine capsules.Suitable carriers for the production of solutions and syrups are, forexample, water, polyols, sucrose, invert sugar, glucose and the like.Suitable carriers for suppositories are, for example, natural orhardened oils, waxes, fats, semi-liquid or liquid polyols and the like.

The pharmaceutical preparations can also contain preservatives,solubilizers, stabilizers, wetting agents, emulsifiers, sweeteners,colorants, flavorants, salts for varying the osmotic pressure, buffers,masking agents or antioxidants. They can also contain therapeuticallyvaluable substances other than the compounds of Formula I.

Medicaments which contain a compound of Formula I with a compatiblepharmaceutical carrier material are also an object of the presentinvention, as is a process for the production of such medicaments whichcomprises bringing one or more of these compounds or salts and, ifdesired, one or more other therapeutically valuable substances into agalenical administration form together with a compatible pharmaceuticalcarrier.

As mentioned earlier, the compounds of Formula I can be used inaccordance with the invention as therapeutically active substances,especially as anti-inflammatory agents or for the prevention of graftrejection following transplant surgery. The dosage can vary within widelimits and will, of course, be fitted to the individual requirements ineach particular case. In general, in the case of administration toadults a convenient daily dosage should be about 0.1 mg/kg to about 100mg/kg, preferably about 0.5 mg/kg to about 5 mg/kg. The daily dosage maybe administered as a single dose or in divided doses and, in addition,the upper dosage limit referred to earlier may be exceeded when this isfound to be indicated.

Finally, the use of compounds of Formula I for the production ofmedicaments, especially in the treatment or prophylaxis of inflammatory,immunological, oncological, bronchopulmonary, dermatological andcardiovascular disorders, in the treatment of asthma, central nervoussystem disorders or diabetic complications or for the prevention ofgraft rejection following transplant surgery, is also an object of theinvention.

Methods of Using the Compounds and Compositions

Compounds of Formula I are useful for, but not limited to, the treatmentof any disorder or disease state in a human, or other mammal, which isexacerbated or caused by excessive or unregulated TNF or p38 kinaseproduction by such mammal. Accordingly, the present invention provides amethod of treating a cytokine-mediated disease which comprisesadministering an effective cytokine-interfering amount of a compound ofFormula I, or a pharmaceutically acceptable salt or tautomer thereof.

Compounds of Formula I are useful for, but not limited to, the treatmentof inflammation in a subject, and for use as antipyretics for thetreatment of fever. Compounds of the invention would be useful to treatarthritis, including but not limited to, rheumatoid arthritis,spondyloarthropathies, gouty arthritis, osteoarhritis, psoriaticarthritis, ankylosing spondylitis, systemic lupus erythematosus andjuvenile arthritis, osteoarthritis, gouty arthritis and other arthriticconditions. Such compounds would be useful for the treatment ofpulmonary disorders or lung inflammation, including adult respiratorydistress syndrome, pulmonary sarcoidosis, asthma, silicosis, and chronicpulmonary inflammatory disease. The compounds are also useful for thetreatment of viral and bacterial infections, including sepsis, septicshock, gram negative sepsis, malaria, meningitis, cachexia secondary toinfection or malignancy, cachexia secondary to acquired immunedeficiency syndrome (AIDS), AIDS, ARC (AIDS related complex), pneumonia,and herpes virus. The compounds are also useful for the treatment ofbone resorption diseases, such as osteoporosis, endotoxic shock, toxicshock syndrome, reperfusion injury, autoimmune disease including graftvs. host reaction and allograft rejections, cardiovascular diseasesincluding atherosclerosis, thrombosis, congestive heart failure, andcardiac reperfusion-injury, renal reperfusion injury, liver disease andnephritis, and myalgias due to infection.

The compounds are also useful for the treatment of Alzheimer's disease,influenza, multiple sclerosis, cancer, diabetes, systemic lupuserthrematosis (SLE), skin-related conditions such as psoriasis, eczema,burns, dermatitis, keloid formation, and scar tissue formation. Inaddition, compounds of the invention are useful in treatinggastrointestinal conditions such as inflammatory bowel disease, Crohn'sdisease, gastritis, irritable bowel syndrome and ulcerative colitis. Thecompounds are also useful in the treatment of ophthalmic diseases, suchas retinitis, retinopathies, uveitis, ocular photophobia, and of acuteinjury to the eye tissue. The compounds can also be used in treatingangiogenesis, including neoplasia; metastasis; opthalmologicalconditions such as corneal graft rejection, ocular neovascularization,retinal neovascularization including neovascularization following injuryor infection, diabetic retinopathy, retrolental fibroplasia andneovascular glaucoma; ulcerative diseases such as gastric ulcer;pathological, but non-malignant, conditions such as hemangiomas,including infantile hemangiomas, angiofibroma of the nasopharynx andavascular necrosis of bone; diabetic nephropathy and cardiomyopathy; anddisorders of the female reproductive system such as endometriosis. Thecompounds can further be used for preventing the production ofcyclooxygenase-2 and have analgesic properties. Therefore, Compounds ofFormula I are useful for treatment of pain.

Other uses for Compounds of Formula I include treatment of HCV, severeasthma, psoriasis, chronic obstructive pulmonary disease (COPD), andother diseases that can be treated with an anti-TNF compound.

Besides being useful for human treatment, these compounds are alsouseful for veterinary treatment of companion animals, exotic animals andfarm animals, including mammals, rodents, and the like. More preferredanimals include horses, dogs, and cats.

The present compounds can also be used in co-therapies, partially orcompletely, in place of other conventional antiinflammatories, such astogether with steroids, cyclooxygenase-2 inhibitors, NSAIDs, DMARDS,immunosuppressive agents, 5-lipoxygenase inhibitors, LTB₄ antagonistsand LTA₄ hydrolase inhibitors.

As used herein, the term “TNF mediated disorder” refers to any and alldisorders and disease states in which TNF plays a role, either bycontrol of TNF itself, or by TNF causing another monokine to bereleased, such as but not limited to IL-1, IL-6 or IL-8. A disease statein which, for instance, IL-1 is a major component, and whose productionor action, is exacerbated or secreted in response to TNF, wouldtherefore be considered a disorder mediated by TNF.

As used herein, the term “p38 mediated disorder” refers to any and alldisorders and disease states in which p38 plays a role, either bycontrol of p38 itself, or by p38 causing another factor to be released,such as but not limited to IL-1, IL-6 or IL-8. A disease state in which,for instance, IL-1 is a major component, and whose production or action,is exacerbated or secreted in response to p38, would therefore beconsidered a disorder mediated by p38.

As TNF-β has close structural homology with TNF-α (also known ascachectin), and since each induces similar biologic responses and bindsto the same cellular receptor, the synthesis of both TNF-α and TNF-β areinhibited by the compounds of the present invention and thus are hereinreferred to collectively as “TNF” unless specifically delineatedotherwise.

EXAMPLES

Additional objects, advantages, and novel features of this inventionwill become apparent to those skilled in the art upon examination of thefollowing illustrative examples thereof, which are not intended to belimiting.

Unless otherwise stated, all temperatures including melting points(i.e., MP) are in degrees celsius (° C.).

Example 1 Preparation of6-(2,4-difluoro-phenoxy)-2-((S)-(+)-2-hydroxy-1-methyl-ethylamino)-8-((S)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pryimidin-7-one

Step A: Preparation ofethyl-4-((S)-2-Hydroxy-propylamino)-2-methylsulfanyl-pyrimidine-5-carboxylate

To a solution of ethyl 4-chloro-2-methylthiopyrimidine-5-carboxylate(Aldrich, 65 g, 280 mmol) in 500 mL of tetrahydrofuran at 0° C. wasadded triethylamine (140 mL 1000 mmol) and (S)-1-amino-2-propanol (21 g,280 mmol). After stirring for 4 hours, water (200 mL) was added and thephases were separated. The aqueous layer was extracted withdichloromethane. The organic phase was concentrated and the residue wasdissolved up with the dichloromethane and washed with brine and driedover magnesium sulfate. Filtered and the filtrate was evaporated underreduced pressure to give 77 g of the ethyl4-(S)-2-hydroxy-propylamino)-2-methylsulfanyl pyrimidine-5-carboxylateas a white solid.

Step B: Preparation of 4-((S)-2-Hydroxy-propylamino)-2-methylsulfanylpyrimidine-5-methanol

Lithium aluminum hydride (5.7 g, 150 mmol) was stirred in drytetrahydrofuran (500 mL) at 5° C. and treated dropwise with a solutionof ethyl 4-((S)-2-hydroxy-propylamino)-2-methylsulfanylpyrimidine-5-carboxylate (27 g, 100 mmol) in dry tetrahydrofuran (450mL). The reaction mixture was stirred for 15 minutes and then water (18mL) was added dropwise with caution. The reaction was stirred for 30minutes and then an aqueous solution of sodium hydroxide (15%, 8.5 mL)was added dropwise, followed by water (25.5 mL). The resultingsuspension was stirred for 17 hours at room temperature and thenfiltered. The filter residue was washed with isopropanol (2×, 100 mL)and the combined filtrate and washings were evaporated under reducedpressure to give 25.8 g 4-((S)-2-hydroxy-propylamino)-2-methylsulfanylpyrimidine-5-methanol.

Step C: Preparation of4-((S)-2-Hydroxy-propylamino)-2-methylsulfanyl-pyrimidine-5-carbaldehyde

4-((S)-2-hydroxy-propylamino)-2-methylsulfanyl pyrimidine-5-methanol (26g, 100 mmol) and 1 L of dichloromethane were combined with stirring andtreated with manganese dioxide (102 g, 1 mol). The resulting suspensionwas stirred for 24 hours and then filtered through celite. The filterresidue was washed with dichloromethane (100 mL) and the combinedfiltrate and washings were evaporated under reduced pressure to give16.5 g of the4-((S)-2-hydroxy-propylamino)-2-methylsulfanyl-pyrimidine-5-carbaldehydeas a white solid.

Sulfone

Step A: Preparation of6-(2,4-Difluorophenoxy)-8-((S)-2-hydroxypropyl)-2-methylsulfanyl-8H-pyrido[2,3-d]pyrimidin-7-one

To a mixture of4-((S)-2-hydroxy-propylamino)-2-methylsulfanyl-pyrimidine-5-carbaldehyde(16.5 g, 73 mmol) and (2,4-difluorophenoxy)acetic acid methyl ester(29.4 g, 145 mmol) in anhydrous dimethyl formamide (300 mL) was addedpotassium carbonate (30 g, 218 mmol). The reaction mixture was heated to60° C. and after 18 hours, the reaction mixture was cooled and thedimethylformamide was distilled off under vacuum. Crude residuesuspended in water (300 mL) and extracted with dichloromethane, washedwith brine and dried over magnesium sulfate. Filtered and concentratedunder vacuum to give 41 g crude material which was chromatographed onsilica gel column eluding with 1% methanol in dichloromethane to give 30g6-(2,4-difluorophenoxy)-8-((S)-2-hydroxypropyl)-2-methylsulfanyl-8H-pyrido[2,3-d]pyrimidin-7-one(mass spec M+1=274).

Step B: Preparation of6-(2,4-Difluorophenoxy)-8-((S)-2-hydroxypropyl)-2-methanesulfonyl-8H-pyrido[2,3-d]pyrimidin-7-one

To a dichloromethane (500 mL) solution of6-(2,4-difluorophenoxy)-8-((S)-2-hydroxypropyl)-2-methylsulfanyl-8H-pyrido[2,3-d]pyrimidin-7-one(29.7 g, 108 mmol) at 5° C. was added portionwise m-chloroperbenzoicacid (55 g, 240 mmol) and stirred for 24 hours. Reaction mixture washedwith aqueous sodium sulfite, aqueous sodium bicarbonate and dried overmagnesium sulfate. Filtered and evaporated to give 24 g6-(2,4-difluorophenoxy)-8-((S)-2-hydroxypropyl)-2-methanesulfonyl-8H-pyrido[2,3-d]pyrimidin-7-one(mass spec M+1=412).

Step C: Preparation of6-(2,4-Difluoro-phenoxy)-2-((S)-2-hydroxy-1-methyl-ethylamino)-8-((S)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7-one

To a tetrahydrofuran (5 mL) solution of6-(2,4-difluorophenoxy)-8-((S)-2-hydroxypropyl)-2-methanesulfonyl-8H-pyrido[2,3-d]pyrimidin-7-one(400 mg, 1 mmol) was added (S)-2-amino-1-propanol (0.38 mL, 5 mmol) andstirred overnight at room temperature. Concentrated under vacuum andchromatographed on silica gel eluding with 2% methanol indichloromethane and converted to the hydrochloride salt to give 320 mg6-(2,4-difluoro-phenoxy)-2-((S)-(+)-2-hydroxy-1-methyl-ethylamino)-8-((S)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7-one(mass spec. M+1=407, MP=175.1-179.1° C.).

Example 2 Preparation of6-(2,4-Difluoro-phenoxy)-2-(R)-(−)-2-hydroxy-1-methyl-ethylamino)-8-((S)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7-one

To a tetrahydrofuran (5 mL) solution of6-(2,4-difluorophenoxy)-8-((S)-2-hydroxypropyl)-2-methanesulfonyl-8H-pyrido[2,3-d]pyrimidin-7-one(400 mg, 1 mmol) was added (R)-2-amino-1-propanol (0.38 mL, 5 mmol) andstirred overnight at room temperature. Concentrated under vacuum andchromatographed on silica gel eluding with 2% methanol indichloromethane and converted to the hydrochloride salt to give 370 mg6-(2,4-difluoro-phenoxy)-2-((R)-2-hydroxy-1-methyl-ethylamino)-8-((S)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7-one(mass spec. M+1=407, MP=174.9-178.1° C.).

Example 3 Preparation of6-(2,4-Difluorophenoxy)-2-(2-hydroxy-1,1-dimethyl-ethylamino)-8-((S)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7-one

To a tetrahydrofuran (10 mL) solution of6-(2,4-difluorophenoxy)-8-((S)-2-hydroxypropyl)-2-methanesulfonyl-8H-pyrido[2,3-d]pyrimidin-7-one(717 mg, 1.74 mmol) was added 2-amino-2-methyl-1-propanol (1.55 g, 17.43mmol) and stirred at room temperature overnight, concentrated undervacuum and chromatographed on silica gel eluting with 4% methanol indichloromethane to give, after converting to hydrochloride salt, 291 mgof6-(2,4-difluorophenoxy)-2-(2-hydroxy-1,1-dimethyl-ethylamino)-8-((S)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7-oneas a white solid (mass spec M+1=421, MP=187.4-189.9° C.).

Example 4 Preparation of6-(2,4-difluoro-phenoxy)-2-((S)-2-hydroxy-1-methyl-ethylamino)-8-((R)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7-one

Step A: Preparation ofethyl-4-((R)-2-Hydroxy-pro-ylamino)-2-methylsulfanyl-pyrimidine-5-carboxylate

To a solution of ethyl 4-chloro-2-methylthiopyrimidine-5-carboxylate(Aldrich, 62.6 g, 269 mmol) in 1 L of THF at 0° C. was addedtriethylamine (135 mL 1000 mmol) and (R)-1-amino-2-propanol (30 g, 400mmol). After stirring for 4 hours, evaporated under reduced pressure togive 66.6 g of the ethyl 4-(R)-2-hydroxy-propylamino)-2-methylsulfanylpyrimidine-5-carboxylate as a white solid.

Step B: Preparation of 4-((R)-2-Hydroxy-propylamino)-2-methylsulfanylpyrimidine-5-methanol

Lithium aluminum hydride (14 g, 368 mmol) was stirred in dry THF (500mL) at 5° C. and treated dropwise with a solution of ethyl4-(R)-2-hydroxy-propylamino)-2-methylsulfanyl pyrimidine-5-carboxylate(66.6 g, 246 mmol) in dry THF (150 mL). The reaction mixture was stirredfor 15 min and then water (18 mL) was added dropwise with caution. Thereaction was stirred for 30 min and then an aqueous solution of sodiumhydroxide (15%, 8.5 mL) was added dropwise, followed by water (25.5 mL).The resulting suspension was stirred for 17 hours at RT and thenfiltered. The filter residue was washed with isopropanol (2×, 100 mL)and the combined filtrate and washings were evaporated under reducedpressure to give 58.6 g 4-(R)-2-hydroxy-propylamino)-2-methylsulfanylpyrimidine-5-methanol.

Step C: Preparation of4-((R)-2-Hydroxy-propylamino)-2-methylsulfanyl-pyrimidine-5-carbaldehyde

4-(R)-2-Hydroxy-propylamino)-2-methylsulfanyl pyrimidine-5-methanol(58.6 g, 256 mmol) and 1 L of dichloromethane were combined withstirring and treated with manganese dioxide (222 g, 2560 mol). Theresulting suspension was stirred for 24 hours and then filtered throughcelite. The filter residue was washed with dichloromethane (100 mL) andthe combined filtrate and washings were evaporated under reducedpressure to give 34 g of the4-((R)-2-hydroxy-propylamino)-2-methylsulfanyl-pyrimidine-5-carbaldehydeas a white solid.

Sulfone

Step A: Preparation of6-(2,4-Difluorophenoxy)-8-((R)-2-hydroxypropyl)-2-methylsulfanyl-8H-pyrido[2,3-d]pyrimidin-7-one

To a mixture of4-((R)-2-hydroxy-propylamino)-2-methylsulfanyl-pyrimidine-5-carbaldehyde(17.7 g, 78 mmol) and (2,4-difluorophenoxy)acetic acid methyl ester(31.6 g, 156 mmol) in anhydrous dimethyl formamide (300 mL) was addedpotassium carbonate (30 g, 218 mmol). The reaction mixture was heated to60° C. and after 18 hours, reaction mixture was cooled and DMF wasdistilled off. Residue suspended in water (300 mL) and extracted withdichloromethane, washed with brine and dried over magnesium sulfate.Filtered and concentrated under vacuum to give 29.5 g crude materialwhich was chromatographed on silica gel column eluding with 1% methanolin dichloromethane to give 17.5 g6-(2,4-difluorophenoxy)-8-((R)-2-hydroxypropyl)-2-methylsulfanyl-8H-pyrido[2,3-d]pyrimidin-7-one(mass spec M+1=274).

Step B: Preparation of6-(2,4-Difluorophenoxy)-8-((R)-2-hydroxypropyl)-2-methanesulfonyl-8H-pyrido[2,3-d]pryimidin-7-one

To a dichloromethane (200 mL) solution of6-(2,4-difluorophenoxy)-8-((R)-2-hydroxypropyl)-2-methylsulfanyl-8H-pyrido[2,3-d]pyrimidin-7-one(9.38 g, 24.7 mmol) at 5° C. was added portion wise m-chloroperbenzoic(12.5 g, 54 mmol) acid and stirred for 24 hours. Reaction mixture washedwith aqueous sodium sulfite, aqueous sodium bicarbonate and dried overmagnesium sulfate. Filtered and evaporated to give 10.7 g6-(2,4-difluorophenoxy)-8-((R)-2-hydroxypropyl)-2-methanesulfonyl-8H-pyrido[2,3-d]pyrimidin-7-one(mass spec M+1=412).

Step C: Preparation of6-(2,4-Difluoro-phenoxy)-2-((S)-2-hydroxy-1-methyl-ethylamino)-8-((R)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7-one

To a THF (5 mL) solution of6-(2,4-difluorophenoxy)-8-((R)-2-hydroxypropyl)-2-methanesulfonyl-8H-pyrido[2,3-d]pyrimidin-7-one(615 mg, 1.5 mmol) was added (S)-2-amino-1-propanol (1.2 mL, 15 mmol)and stirred overnight at RT. Concentrated under vacuum andchromatographed on silica gel eluding with 2% methanol indichloromethane and converted to the hydrochloride salt to give 295 mg6-(2,4-difluoro-phenoxy)-2-((S)-2-hydroxy-1-methyl-ethylamino)-8-((R)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7-one(mass spec. M+1=407, MP=186.0-189.1° C.).

Example 5 Preparation of6-(2,4-Difluoro-phenoxy)-2-(R)-2-hydroxy-1-methyl-ethylamino)-8-((R)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7-one

To a tetrahydrofuran (5 mL) solution of6-(2,4-difluorophenoxy)-8-((R)-2-hydroxypropyl)-2-methanesulfonyl-8H-pyrido[2,3-d]pyrimidin-7-one(400 mg, 1 mmol) was added (R)-2-amino-1-propanol (0.38 mL, 5 mmol) andstirred overnight at room temperature. Concentrated under vacuum andchromatographed on silica gel eluding with 2% methanol indichloromethane and converted to the hydrochloride salt to give 350 mg6-(2,4-difluoro-phenoxy)-2-((R)-2-hydroxy-1-methyl-ethylamino)-8-((R)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7-one(mass spec. M+1=407, MP=181.5-184.4° C.).

Example 6 Preparation of6-(2,4-Difluorophenoxy)-2-(2-hydroxy-1,1-dimethyl-ethylamino)-8-((R)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7-one

A mixture of6-(2,4-difluorophenoxy)-8-((R)-2-hydroxypropyl)-2-methanesulfonyl-8H-pyrido[2,3-d]pyrimidin-7-one(886 mg, 2.15 mmol) and 2-amino-2-methyl-1-propanol (5.15 g, 58 mmol)was heated at 60° C. under nitrogen for 2 hours. Cooled andchromatographed on silica gel eluding with 2% methanol indichloromethane to give after converting to hydrochloride salt 385 mg6-(2,4-Difluorophenoxy)-2-(2-hydroxy-1,1-dimethyl-ethylamino)-8-((R)-2-hydroxy-propyl)-8H-pyrido[2,3-d]pyrimidin-7-one(mass spec. M+1=421, MP=182.0-183.9° C.).

Example 7 Preparation of6-(2,4-Difluorophenoxy)-2-((S)-2-hydroxy-1-methyl-ethylamino)-8-(2-oxopropyl)-8H-pyrido[2,3-d]pyrimidin-7-one

Step A: Preparation of6-(2,4-Difluorophenoxy)-2-methanesulfonyl-8-(2-oxopropyl)-8H-pyrido[2,3-d]pyrimidin-7-one

To a dichloromethane (100 mL) solution of oxalyl chloride (1.05 mL, 12mmol) at −60° C. was added dimethyl sulfoxide (1.7 mL, 24 mmol) and6-(2,4-difluorophenoxy)-8-(2-hydroxypropyl)-2-methanesulfonyl-8H-pyrido[2,3-d]pyrimidin-7-one(4.12 g, 10 mmol). To this mixture was added triethylamine (7 mL, 50mmol) and stirred overnight. Added water (100 mL) and extracted withdichloromethane, washed with brine and dried over magnesium sulfate.Filter and concentrated under vacuum, chromatographed on silica geleluding with 2% methanol in dichloromethane to give 1.0 g6-(2,4-fluorophenoxy)-2-methanesulfonyl-8-(2-oxopropyl)-8H-pyrido[2,3-d]pyrimidin-7-one(mass Spec. M+1=410)

Step B: Preparation of6-(2,4-Difluorophenoxy)-2-((S)-2-hydroxy-1-methyl-ethylamino)-8-(2-oxopropyl)-8H-pyrido[2,3-d]pyrimidin-7-one

To a THF suspension of6-(2,4-difluorophenoxy)-2-methanesulfonyl-8-(2-oxopropyl)-8H-pyrido[2,3-d]pyrimidin-7-one(412 mg, 1 mmol) was added (S)-2-amino-1-propanol (0.39 mL, 5 mmol) atRT and stirred overnight. Concentrated under vacuum and chromatographedon silica gel eluding with 2% methanol in dichloromethane to give afterconversion to the hydrochloride salt 330 mg6-(2,4-Difluorophenoxy)-2-((S)-2-hydroxy-1-methyl-ethylamino)-8-(2-oxopropyl)-8H-pyrido[2,3-d]pyrimidin-7-one(Mass spec. M+1=405, MP=207.9-214.6° C.).

Example 8 Preparation of6-(2,4-Difluorophenoxy)-2-((R)-2-hydroxy-1-methyl-ethylamino)-8-(2-oxopropyl)-8H-pyrido[2,3-d]pyrimidin-7-one

To a tetrahydrofuran (10 mL) suspension of6-(2,4-difluorophenoxy)-2-methanesulfonyl-8-(2-oxopropyl)-8H-pyrido[2,3-d]pyrimidin-7-one(417 mg, 1 mmol) was added (R)-(−)-2-amino-1-propanol (0.40 mL, 5 mmol)at room temperature and stirred overnight. Concentrated under vacuum andchromatographed on silica gel eluding with 2% methanol indichloromethane to give after conversion to the hydrochloride salt 330mg6-(2,4-Difluorophenoxy)-2-((R)-2-hydroxy-1-methyl-ethylamino)-8-(2-oxopropyl)-8H-pyrido[2,3-d]pyrimidin-7-one(Mass spec. M+1=405, MP=207.8-216.4° C.).

Example 9

In Vitro Assay

This example illustrates a p38 (MAP) kinase in vitro assay useful forevaluating the compounds of the present invention.

The p38 MAP kinase inhibitory activity of compounds of this invention invitro was determined by measuring the transfer of the γ-phosphate fromγ-³³P-ATP by p-38 kinase to Myelin Basic Protein (MBP), using a minormodification of the method described in Ahn, et al., J. Biol. Chem.266:4220-4227 (1991).

The phosphorylated form of the recombinant p38 MAP kinase wasco-expressed with SEK-1 and MEKK in E. Coli (see, Khokhlatchev, et al.,J. Biol. Chem. 272:11057-11062 (1997)) and then purified by affinitychromatography using a Nickel column.

The phosphorylated p38 MAP kinase was diluted in kinase buffer (20 mM3-(N-morpholino)propanesulfonic acid, pH 7.2, 25 mM β-glycerolphosphate, 5 mM ethylene glycol-bis(beta-aminoethylether)-N,N,N′,N′-tetraacetic acid, 1 mM sodium ortho-vanadate, 1 mMdithiothreitol, 40 mM magnesium chloride). Test compound dissolved inDMSO or only DMSO (control) was added and the samples were incubated for10 min at 30° C. The kinase reaction was initiated by the addition of asubstrate cocktail containing MBP and γ-³³P-ATP. After incubating for anadditional 20 min at 30° C., the reaction was terminated by adding 0.75%phosphoric acid. The phosphorylated MBP was then separated from theresidual γ-³³P-ATP using a phosphocellulose membrane (Millipore,Bedford, Mass.) and quantitated using a scintillation counter (Packard,Meriden, Conn.).

Using the above assay, compounds of the invention were shown to beinhibitors of p38 MAP kinase. The compounds of the invention exhibitedp38 IC₅₀ values in the range of from less than 0.001 to 0.1 μM. Forexample,6-(2,4-difluoro-phenoxy)-8-(3-hydroxy-propyl)-2-(tetrahydro-pyran-4-ylamino)-8H-pyrido[2,3-d]pyrimidin-7-oneshowed an IC₅₀ of 0.0008 μM using the above assay.

Example 10

In Vitro Assay

This example illustrates a human whole blood (HWB) in vitro assay (i.eLPS-induced IL-1β production in undiluted human whole blood viainhibition of p38 MAP kinase) for evaluating the compounds of thepresent invention and the comparative results of corresponding alkylanalogs.

LPS (lipopolysaccharide) treatment of human whole blood induces IL-1β(Interleukin-1β) production that can be measured by an IL-1β specificELISA. Human whole blood was pre-incubated with the indicatedconcentrations of a compound of the present invention in 0.5% DMSO(final concentration) for 30 min at 37° C. Samples were stimulated with0.5 μg/mL of lipopolysaccharide (LPS, from Sigma) (final concentration)for 18 hours to induce the synthesis and secretion of IL-1β which wasmeasured using an IL-1β ELISA.

Solution Preparation

Compound Solutions

A stock solution of 6 mM in DMSO (from Sigma) was prepared by dissolvingthe compound in DMSO in 449 μL of DMSO. From the 6 mM stock solution,six subsequent half-log serial dilutions in DMSO were performed to givethe following concentrations: 1.9 mM, 600, 190, 60, 19, and 6 μM. Labeltubes 1-7. The 6 mM stock solution in tube 1. 216 μL DMSO was placed ineach of tubes 2-7. From tube 1,100 μL was transferred to tube 2. Tube 2was vortexed and 100 μL was transferred from tube 2 to tube 3. Thisprocess was repeated to tube 7.

Using the serial dilutions of the compound in DMSO prepared above, anadditional dilution 1/20 (10 μL into 190 μL RPMI 1640 medium, fromGibco-BRL) was performed to give a final compound concentration curve of30, 10, 2.9, 1, 0.3, 0.1, 0.03 μM.

LPS Solutions

Reconstitution of LPS: in the 10 mg. vial of LPS, 10 mL 1×Phosphate-Buffered Saline (i.e., 1×PBS, from Gibco-BRL) was added, mixedwell and transferred to a 50 mL tube. Another 10 mL was added to the LPSvial, followed by rinsing, and this rinse was added to the 50 mL tubeand mixed will. The solution was filtered and sterilized, and aliquottedin desired amounts (100 μL aliquot was sufficient for 4 plates). Thisyielded a 0.5 mg/mL stock which was diluted 1/100 for use in protocol.Just prior to use, the LPS stock was diluted 1/100 (100 μL in 10 mL ofRPMI)

Assay Procedure

The Assay was performed in a 96 well U bottom plate (from Costar). Twocontrols were included in each assay, plus and minus LPS in the absenceof compound. All samples and controls were performed in triplicate.

Human blood (from donors who had received no medication for at least 14days, no alcohol for 48 hours) was collected into siliconizedvacutainers containing heparin (19 units/ml). A 25 μL aliquot of 5% DMSOin RPMI 1640 was added to control wells (plus and minus LPS controls).25 mL aliquots of each compound concentration prepared above weredispensed to designated wells. 200 μL of human whole blood was added toeach well and incubated at 37° C. and 5% CO₂ for 30 minutes. 25 μL ofdiluted LPS was dispensed to all wells except minus LPS control wells.25 μL of RPMI was added to minus LPS control wells.

The plates were incubated at 37° C. and 5% CO₂ for 18 hours. Afterincubation, plates were centrifuged at 400×g to pellet cells and collectplasma, taking care not to disturb the pellet. The plasma wastransferred to a new 96 well polypropylene plate. ELISA was performedimmediately and remaining plasma was stored at −20° C., in case neededfor re-testing.

ELISA Protocol

The IL-1β ELISA used two anti-IL-1β monoclonal antibodies: ILβ1-H6 (1mg/mL) and ILβ1-H67 (2.71 mg/mL).

Materials

-   -   Recombinant human IL-1β (rhuIL-1β, 2.5 μg/mL) was obtained from        R&D Systems.

Phosphate-Buffered Saline-Dulbecco's (1×PBS) was obtained fromGibco-BRL.

Phosphate-Buffered Saline (10×PBS) was obtained from Gibco-BRL.

Dulbeccos' modification without calcium and magnesium, pH 7.2.

Unopened bottles were stored at Room Temperature.

ELISA Incubation Buffer (EIB)

-   -   0.1% BSA/PBS    -   1 g Bovine serum albumin (BSA)    -   100 mL of 10×PBS    -   Add deionized water to 1 liter and store at 4° C.

ELISA Wash Buffer (EWB)

-   -   0.05% Tween/PBS    -   0.5 mL Tween 20    -   100 mL 10×PBS    -   Add deionized water to 1 liter and store at 4° C.

Blocking Buffer—3% Nonfat-dry milk/PBS

-   -   15 g nonfat-dry milk powder (Carnation)    -   50 mL 10×PBS    -   Add distilled water to 500 mL and store at 4° C.

Peroxidase Conjugated Streptavidin (from Pharmingen)

-   -   Dilute approximately 1:3000 (10 μL/30 mL) in EWB buffer.

0.1 M Citrate Buffer, pH 4.5

-   -   9.6 g. citric acid (MW 192.1, from Sigma)    -   14.7 g. tri-sodium citrate (MW 294.1, from Sigma)    -   Adjust to pH 4.5 using NaOH and add distilled water to 500 mL.        Store at 4° C.

OPD Substrate solution

-   -   1 mg/mL OPD/0.03% H₂O₂/citrate buffer    -   1 tablet o-phenylenediamine (OPD, from Zymed)    -   12 μL of 30% hydrogen peroxide    -   12 mL of 0.1 M citrate buffer        Preparation of Standards (Prepare Fresh Just Prior to Placing on        Plate).

A stock solution of rhuIL-1β (2.5 μg/mL) was used to construct astandard curve. The concentrations for the curve are: 12500, 4167, 1389,463, 154, 51 and 17 pg/mL. Tubes were labeled 1-8. rhuIL-1β stocksolution was diluted 1/500 (3 μL of stock+597 μL of EWB) in tube 1. 400μL EWB was dispensed in tubes 2-8. From tube 1, 200 μL was transferredto tube 2 and vortexed. 200 μL was transferred from tube 2 to tube 3.Repeat this process to tube 7. Tube 8 was used as the ELISA assay blank.

Plasma samples were diluted 1:4 in EWB (20 μL of plasma+60 μL of EWB).

Preparation of Antibody Solutions.

Antibody ILβ1-H6 was diluted 1/100 in 1×PBS to generate 10 μg/mLsolution. Per plate, 50 μL of antibody was diluted in 5 mL PBS. AntibodyILβ1-H67 was diluted 1/100 in EWB to generate 2 μg/mL solution. Perplate, 3.69 μL of antibody was diluted in 5 mL EIB.

Procedure

96-well EIA plates were coated with 50 μL per well of antibody ILβ1-H6(10 μg/mL), shaken gently to clear any air bubbles, and sealed withplate sealer and incubate in a humidified chamber overnight at 4° C. Theplates were emptied and tapped dry on a lint free paper towel.Non-specific binding sites were blocked with 175 μL per well of BlockingBuffer for 1-2 hours at room temperature. The plates were washed oncewith EWB (i.e., empty plate, filled with 150 μL EWB, emptied and tappeddry on a lint free paper towel).

Triplicate 25 μL aliquots of standard were added to appropriate wells.(Each plate has its own standard curve.). A 25 μL aliquot of dilutedplasma was added to the appropriate well. To all wells, 25 μL ofbiotinylated monoclonal antibody ILβ1-H67 (2 μg/mL) was added. Theplates were sealed with plate sealer and incubated for 2 hours at roomtemperature (or overnight at 4° C.) while gently shaking (BellcoMini-Orbital Shaker, setting 3.5). After incubation, plates were washed3× with EWB (as described above). A 50 μL aliquot ofperoxidase-streptavidin, diluted 1:3000 in EIB, was added to each well.Plates were sealed with plate sealer, incubated plates for 1 hour atroom temperature while shaking, and washed 3× as described above.

An OPD tablet was dissolved in citrate buffer, (1 tablet/12 mL citratebuffer), and 12 μL of 30% H₂O₂ was added to OPD/citrate buffer). 50 μLof OPD substrate solution was dispensed to each well, and plates wereincubated in the dark for 30 minutes at room temperature for colordevelopment. Plates were read at dual wavelength: Sample filter=450nm/Reference filter=650 nm. The values from the samples containing thestandard were used to graph a standard curve (absorbance vs.concentration) used to determine concentrations of unknown samples.

Statistical Method

If the concentration-inhibition curve does not include points on eitherside of 50%, then the IC₅₀ was reported as >highest concentration or<lowest concentration. Otherwise, if the number of concentration was ≧5,the data was fitted to the following 2-parameter model to estimate anIC₅₀:

${{Mean}\mspace{14mu}\%\mspace{14mu}{Inhibition}} = \frac{100}{1 + \left( \frac{{IC}_{50}}{Conc} \right)^{n}}$

This model assumes the minimum and maximum response were 0% and 100%,respectively, and estimates the IC₅₀ and the slope parameter. If thenonlinear regression fails or if the number of concentrations tested was<5, linear regression was used to estimate the IC₅₀ using the 2 pointsthat flank 50%.

If linear regression was used to estimate the IC₅₀, this was output inthe assay notes, and can also be seen with the presence (nonlinearregression) or absence (linear regression) of the IC₅₀ standard errorand the slope parameter.

Results

Using the above assay, compounds of the invention were shown to inhibitLPS-induced IL-1β production in undiluted human whole blood viainhibition of p38 MAP kinase, which mediates IL-1β production as notedabove. Compounds of the invention exhibited IC₅₀ values for LPS-inducedIL-1β production in undiluted human whole blood in the range of from<0.001 μM to 0.30 μM. For example,6-(2,4-difluoro-phenoxy)-8-((R)-2-hydroxy-propyl)-2-(tetrahydro-pyran-4-ylamino)-8H-pyrido[2,3-d]pyrimidin-7-oneshowed an IC₅₀ of 0.001 μM.

Surprisingly, the inhibition of LPS-induced IL-1β production usingcompounds of the invention wherein R² of formula (I) is hydroxyalkyl oralkoxyalkyl, is substantially greater than results from correspondingcompounds wherein R² is methyl or other alkyl. This unexpected advantageof the invention is illustrated more fully in Table 2, in whichrepresentative compounds of the invention where R² (of formula I) ishydroxyalkyl are compared to the corresponding analogs where R² ismethyl). Compounds in the first or leftmost column of Table 1 areprepared as described in the examples herein, and are also shown inTable 1. Compounds in the second or center column of Table 2 wereprepared according to the procedures reported in WO 02/064594. Thevalues in the third or right-most column correspond to the ratio:(IC₅₀ inhibition of IL-1β production where R²=hydroxyalkyl)/(IC₅₀inhibition of IL-1β production where R²=methyl).

As can be seen from Table 2, compounds wherein R² is hydroxyalkylprovide inhibition of LPS-induced IL-1β production in undiluted humanwhole blood that is on the order of 2.7 to >100 times, i.e., 270%to >10,000%, greater than the corresponding methyl analogs (12=methyl).

TABLE 2 IC₅₀/ IC₅₀ R² = hydroxyalkyl R² = alkyl (methyl) ratio

43.4

27.8

9.9

100

17.9

5.3

17.4

24.6

2.7

200

7.3

11.4

4.7

3.9

6.2

The foregoing discussion of the invention has been presented forpurposes of illustration and description. The foregoing is not intendedto limit the invention to the form or forms disclosed herein. Althoughthe description of the invention has included description of one or moreembodiments and certain variations and modifications, other variationsand modifications are within the scope of the invention, e.g., as may bewithin the skill and knowledge of those in the art, after understandingthe present disclosure. It is intended to obtain rights which includealternative embodiments to the extent permitted, including alternate,interchangeable and/or equivalent structures, functions, ranges or stepsto those claimed, whether or not such alternate, interchangeable and/orequivalent structures, functions, ranges or steps are disclosed herein,and without intending to publicly dedicate any patentable subjectmatter. All publications, patents, and patent applications cited hereinare hereby incorporated by reference in their entirety for all purposes.

1. A compound of the formula:

wherein X¹ is O, S(O)_(n), or C═O; wherein n is 0, 1 or 2; Ar¹ is arylor heteroaryl; R¹ is hydroxyalkyl; and R² is oxoalkyl.
 2. The compoundof claim 1, wherein Ar¹ is aryl.
 3. The compound of claim 1, wherein Ar¹is heteroaryl.
 4. The compound of claim 2, wherein X¹ is O.
 5. Thecompound of claim 4, wherein: R¹ is (R)-2-hydroxy-1-methylethyl or(S)-2-hydroxy-1-methylethyl.
 6. A compound of the formula:

wherein: m is from 0 to 4; R¹ is hydroxyalkyl; R² is oxoalkyl; and eachR³ independently is: alkyl; alkoxy; halo; or haloalkyl.
 7. The compoundof claim 6, wherein R³ is halo.
 8. The compound of claim 6, wherein m isfrom 0 to
 2. 9. The compound of claim 6, wherein n is 2 and R³ is halo.10. The compound of claim 1, wherein said compound is selected from:6-(2,4-Difluoro-phenoxy)-2-((S)-2-hydroxy-1-methyl-ethylamino)-8-(2-oxo-propyl)-8H-pyrido[2,3 -d]pyrimidin-7-one; and6-(2,4-Difluoro-phenoxy)-2-((R)-2-hydroxy-1-methyl-ethylamino)-8-(2-oxo-propyl)-8H-pyrido [2,3 -d]pyrimidin-7-one.
 11. A composition comprising: (a) apharmaceutically acceptable excipient; and (b) a compound of claim 1.12. A compound of the formula:

wherein X¹ is O, S(O)_(n), or C═O; wherein n is 0, 1 or 2; Ar¹ is phenyloptionally substituted one or more times with alkyl, halo, haloalkyl oralkoxy; R¹ is hydroxyalkyl; and R² is oxoalkyl.
 13. The compound ofclaim 12, wherein X¹ is O.
 14. The compound of claim 13, wherein Ar¹ is2,4-disubstituted phenyl.
 15. The compound of claim 14, wherein Ar¹ is2,4-halorophenyl.
 16. The compound of claim 15, wherein Ar¹ is2,4-difluorophenyl.
 17. The compound of claim 16, wherein: R¹ is(R)-2-hydroxy-1-methylethyl or (S)-2-hydroxy-1-methylethyl.